表达IBDV AH1株VP2嵌合法氏囊素三肽基因的重组火鸡疱疹病毒的构建及免疫功能分析  

Construction of recombinant turkey herpesvirus expressing the VP2 protein of infectious bursal disease virus AH1 strain and its immunologic functions

作　　者：潘群兴[1] 毕振威[1] 王永山[1] 欧阳伟[1] 王晓丽[1] 孟凯[1] 夏兴霞[1] 诸玉梅[1] 

机构地区：[1]江苏省农业科学院兽医研究所,农业部兽用生物制品工程技术重点实验室,国家兽用生物制品工程技术研究中心,江苏南京210014

出　　处：《中国兽医科学》2017年第9期1073-1079,共7页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2016YFD0500800);国家自然科学基金项目(31272537,31302070,31572504);江苏省自然科学基金项目(BK20141381,BK20151366);江苏省农业科技自主创新资金项目[CX(16)1052]

摘　　要：为研究传染性法氏囊病(IBD)重组火鸡疱疹病毒(HVT)的构建策略,以HVT FC126基因组非必需区UL44~UL45为插入位点,构建带有大肠杆菌黄嘌呤-鸟嘌呤磷酸转移酶基因序列(gpt)标记的HVT转移载体质粒pUAB-gpt。然后构建表达IBDVAH1株VP2嵌合法氏囊素三肽(BS10)表达盒并克隆入pUAB-gpt,获得转移载体质粒pUAB-Pec-BS10-VP2-Egpt。将该转移载体与HVT DNA共转染鸡胚成纤维细胞(CEF),通过间接免疫荧光试验(IFA)、IBDV抗体夹心ELISA检测、Western-blotting分析,结果显示,获得了表达VP2基因的重组HVT病毒rHVT-Pec-BS10-VP2。将该重组病毒免疫1日龄SPF鸡,免疫后第14天,鸡群可以检测到IBDV抗体。免疫后第21天,脾淋巴细胞增殖活性反应显著。证明IBD重组火鸡疱疹病毒构建成功。Construction strategy and immune functions of recombinant herpesvirus of infectious bursal disease （IBD） were studied.Non essential region UL44-UL45 of the HVT FC126 genome was selected as insertion sites.Using the selection marker Eco-gpt（Xanthine-guanine phosphoribosyl transferase（gpt）,plasmid pUAB-gpt was constructed.Then,BS10-VP2 expression cassette was constructed and cloned into the plasmid pUAB-gpt to obtain the transfer vector of turkey herpesvirus pUAB-Pec-BS10-VP2-Egpt.Finally,transfer vector and total DNA of HVT infected cells were cotransfected into chicken embryo fibroblasts （CEFs）.The recombinant rHVT-Pec-BS10-VP2 expressing the VP2 gene of IBDV was identified by indirectimmunofluorescenceassay （IFA）,IBDVsandwich ELISAand Western-blot.Inaddition,ther HVT-Pec-BS10-VP2 were used to immunize 1-day-old SPF chicken.Antibody titrations were detected on 14 day after immunization.Splenic lymphocyte proliferation activity was remarkable on 21 day after immunization.Theseresults demonstrate that the recombinant HVT expressing the VP2 protein of infectious bursal disease virus is proved successful.

关 键 词：传染性法氏囊病病毒 VP2基因 法氏囊素三肽 重组火鸡疱疹病毒 

分 类 号：S852.659.4[农业科学—基础兽医学]